CN113273605A - Chitosan-silver complex antibacterial gel, preparation method and fresh-keeping application thereof - Google Patents
Chitosan-silver complex antibacterial gel, preparation method and fresh-keeping application thereof Download PDFInfo
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- CN113273605A CN113273605A CN202110386009.0A CN202110386009A CN113273605A CN 113273605 A CN113273605 A CN 113273605A CN 202110386009 A CN202110386009 A CN 202110386009A CN 113273605 A CN113273605 A CN 113273605A
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- 229910052709 silver Inorganic materials 0.000 title claims abstract description 40
- 239000004332 silver Substances 0.000 title claims abstract description 40
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000000017 hydrogel Substances 0.000 claims abstract description 30
- 229920001661 Chitosan Polymers 0.000 claims abstract description 28
- 229910001961 silver nitrate Inorganic materials 0.000 claims abstract description 16
- 238000004321 preservation Methods 0.000 claims abstract description 8
- -1 silver ions Chemical class 0.000 claims abstract description 8
- 239000002131 composite material Substances 0.000 claims abstract 2
- 239000011550 stock solution Substances 0.000 claims description 24
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 241000219094 Vitaceae Species 0.000 claims description 13
- 235000021021 grapes Nutrition 0.000 claims description 13
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 9
- 239000012498 ultrapure water Substances 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 8
- 238000004090 dissolution Methods 0.000 claims description 5
- 239000000243 solution Substances 0.000 claims description 5
- 229960000583 acetic acid Drugs 0.000 claims description 4
- 239000012362 glacial acetic acid Substances 0.000 claims description 4
- 239000011259 mixed solution Substances 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 2
- 238000001338 self-assembly Methods 0.000 claims description 2
- 239000013078 crystal Substances 0.000 claims 1
- 238000000034 method Methods 0.000 claims 1
- 239000000499 gel Substances 0.000 abstract description 12
- 235000009754 Vitis X bourquina Nutrition 0.000 abstract description 7
- 235000012333 Vitis X labruscana Nutrition 0.000 abstract description 7
- 235000014787 Vitis vinifera Nutrition 0.000 abstract description 7
- 230000003385 bacteriostatic effect Effects 0.000 abstract description 4
- 230000002195 synergetic effect Effects 0.000 abstract 2
- 240000006365 Vitis vinifera Species 0.000 abstract 1
- 230000003115 biocidal effect Effects 0.000 abstract 1
- 239000003638 chemical reducing agent Substances 0.000 abstract 1
- 239000002270 dispersing agent Substances 0.000 abstract 1
- 238000002604 ultrasonography Methods 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 7
- 241000219095 Vitis Species 0.000 description 6
- 238000003860 storage Methods 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000012452 mother liquor Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 241000228245 Aspergillus niger Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 108010093096 Immobilized Enzymes Proteins 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 108010081750 Reticulin Proteins 0.000 description 1
- ATJFFYVFTNAWJD-UHFFFAOYSA-N Tin Chemical compound [Sn] ATJFFYVFTNAWJD-UHFFFAOYSA-N 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000009920 food preservation Methods 0.000 description 1
- 239000005452 food preservative Substances 0.000 description 1
- 235000019249 food preservative Nutrition 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229920001477 hydrophilic polymer Polymers 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/16—Coating with a protective layer; Compositions or apparatus therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/154—Organic compounds; Microorganisms; Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/157—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Inorganic Chemistry (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Storage Of Fruits Or Vegetables (AREA)
Abstract
The invention provides chitosan-silver hydrogel with excellent antibacterial property, a preparation method and application thereof, and provides chitosan-silver complex antibacterial gel, and a preparation method and application thereof. The preparation method of the composite hydrogel comprises the following steps: silver nitrate and chitosan are assembled together under the action of ultrasound to form clear and transparent complex hydrogel. In the hydrogel, the synergistic effect of silver ions and chitosan greatly improves the antibacterial and fresh-keeping properties of the hydrogel. The invention has the performances of antibiosis, fresh preservation, easy smearing and the like; the preparation method is simple, the cost is low, commercialization can be realized, the bacteriostatic effect is good, the stability is good, and the like; no reducing agent or dispersing agent is added in the preparation process, so that the preparation method is green and environment-friendly. The chitosan-silver hydrogel disclosed by the invention has the advantages that the antibacterial property is obviously improved through the synergistic effect of the components, and the chitosan-silver hydrogel has a wide application prospect in the field of grape preservation.
Description
Technical Field
The invention belongs to the technical field of food preservation, and particularly relates to a chitosan-silver antibacterial gel for grape preservation, and a preparation method and application thereof.
Background
The grape is easy to damage during storage and transportation due to the fragile surface and high water content, so that the grape is infected and propagated by microorganisms to rot and deteriorate, and the preservation quality and the economic benefit of the grape are seriously influenced. According to statistics, the grape storage and fresh-keeping treatment is poor in China, so that the annual storage loss rate of grapes is 20% -25%, the logistics loss rate is up to 40%, and the logistics loss rate is far lower than that of grapes in developed countries. The storage, transportation and preservation of grapes are becoming more and more prominent, and how to prolong the storage time of grapes, keep the quality of grapes unchanged and inhibit the invasion of microorganisms is a problem to be solved urgently.
The traditional chemical food preservative has been gradually replaced due to the influence on human health and the disadvantages of environmental pollution and the like. The selection of natural, green and safe antistaling agents is widely concerned. Chitosan (CS for short) and its derivatives are used as a natural biological macromolecule, and have good broad-spectrum antibacterial activity, moisture retention property and no toxic or side effect, thus entering the sight of people. However, the pure chitosan has a narrow antibacterial spectrum, the antibacterial effect is not very obvious, and the adsorbability of the chitosan is generally low in coating efficiency, so that the further application of the chitosan is limited. Silver and its compounds are among the most common antimicrobial agents, having a broad spectrum of antimicrobial properties. However, silver ions are easily oxidized by air to affect the bacteriostatic effect, and the excessive intake of silver ions may cause harm to the liver of a human body. Therefore, how to coordinate the two natural antibacterial preservatives and improve the application effect has important significance for the application in the antibacterial and fresh-keeping field.
The polysaccharide hydrogel is mainly a hydrophilic polymer network system, has extremely high water content (generally more than 90% of the weight of the hydrogel is water molecules), and has excellent biocompatibility and biodegradability. Substances encapsulated in the hydrogel can be tightly fixed by the 3D reticular fibers, and air can be isolated, so that the oxidation of the encapsulated substances is avoided, and the hydrogel has an important effect in the researches of immobilized enzyme, cell separation, protein preparation, slow-release medicines and the like.
By utilizing the principle, the chitosan-silver complex hydrogel with double bacteriostatic and fresh-keeping properties is successfully prepared. The preparation process of the invention has simple operation and low cost, and is suitable for large-scale production. Experimental results show that the chitosan-silver complex hydrogel is expected to have good application in the fields of application of antibacterial agents and preservation and the like.
Disclosure of Invention
The invention aims to provide a preparation method and application of chitosan-silver complex hydrogel with excellent antibacterial and freshness-keeping properties. The chitosan-silver gel is clear and transparent hydrogel, has spreadability, can enhance the resistance of grapes to bacteria, and achieves a long-term fresh-keeping effect.
The preparation method of the antibacterial chitosan-silver complex gel comprises the following steps:
(1) preparing a chitosan stock solution with a certain concentration;
(2) preparing a silver nitrate stock solution with a certain concentration;
(3) mixing and shaking up a certain amount of the stock solutions of the components, and carrying out ultrasonic treatment to obtain the chitosan-silver complex hydrogel.
In the step (1) of the preparation method, the solvent of the chitosan stock solution is ultrapure water, the concentration of the chitosan stock solution is 5-20 mg/mL, and glacial acetic acid with the volume ratio of 1% is added to promote the dissolution of chitosan.
In the step (2), the solvent of the silver nitrate stock solution is ultrapure water, the concentration of the silver nitrate stock solution is 0.3 mol/L, and the silver nitrate stock solution is processed in a dark place during preparation.
In the step (3), 100. mu.L of silver nitrate stock solution is added into a 2.0mL screw bottle which is processed in a dark place and quickly shaken up, 800. mu.L of chitosan stock solution and 100. mu.L of sodium hydroxide solution (0.2mol/L) are added to prepare 1.0mL mixed solution, and the mixed solution is quickly shaken up to form the chitosan-silver complex hydrogel.
The chitosan-silver gel used in the invention is obtained by non-covalent bond self-assembly, and the hydrogel can quickly form gel by shaking. In addition, chitosan is combined with silver particles, so that the organic antibacterial property and the freshness retaining property of the hydrogel are enhanced.
Advantageous effects
(1) The chitosan-silver complex gel prepared by the invention has good biocompatibility, is easy to degrade and has no cytotoxicity.
(2) The chitosan-silver complex gel prepared by the invention is clear and transparent hydrogel, is easy to smear and has good moisture retention.
(3) The chitosan and the silver ions added in the invention have broad-spectrum antibacterial property respectively, and the antibacterial property of the combination of the chitosan and the silver ions is obviously improved.
(4) The preparation method is simple, low in cost, capable of being commercialized and suitable for large-scale production.
(5) The chitosan-silver complex gel prepared by the invention has good antibacterial property and freshness retaining property, and has good application prospect in the field of fruit freshness retaining.
Drawings
FIG. 1 shows gel-forming concentrations of chitosan-silver complex hydrogels:
FIG. 2 is a UV spectrum of a chitosan-silver complex in example 1 of the present invention;
FIG. 3 is a Scanning Electron Microscope (SEM) image of a chitosan-silver complex hydrogel;
FIG. 4 is a digital photographic image of the effect of chitosan-silver complexes on the growth of Penicillium;
FIG. 5 is a digital photograph showing the effect of chitosan-silver complex on the growth of Aspergillus niger.
Detailed Description
In order to facilitate understanding of the invention, the invention will be described in more complete detail below with reference to the drawings and specific examples, but the scope of the invention is not limited to the specific examples below.
Example 1
Weighing 2.046g of silver nitrate into a brown light-proof centrifuge tube, adding 20mL of ultrapure water to dissolve the silver nitrate, and preparing 0.6mol/L silver nitrate aqueous solution into 50mL of brown reagent subjected to light-proof treatment by using tinfoil paper to serve as silver nitrate stock solution; 500mg of chitosan is weighed, 100mL of ultrapure water is added to dissolve the chitosan, and glacial acetic acid with the volume ratio of 1% is added to promote the dissolution of the chitosan, so that 5mg/mL of chitosan stock solution is prepared. 160mg of sodium hydroxide was weighed and dissolved in 20mL of ultrapure water to prepare a 0.2mol/mL stock solution of sodium hydroxide.
And adding 100 mu L of silver nitrate stock solution into a 2.0mL screw bottle which is processed in a dark place, quickly shaking up, adding 800 mu L of chitosan stock solution and 100 mu L of sodium hydroxide solution to prepare 2.0mL mixed solution, and quickly shaking up to form clear and transparent chitosan-silver hydrogel.
The antibacterial experiment steps are as follows: weighing 46g of PDA culture medium powder in a beaker, adding ultrapure water into the beaker for dissolution, stirring with a glass rod to promote dissolution, and preparing the PDA culture medium in a conical flask with the volume being constant to 1L by using the ultrapure water.
Taking a proper amount of naturally rotten red globe grapes, adding the red globe grapes into a beaker containing 100mL of sterile water, continuously stirring, taking out the grapes, and filtering and collecting liquid containing fungal spores. The filtrate was regarded as mother liquor of spore suspension, and the mother liquor was diluted to 10-1, 10-2, 10-3, 10-4, 10-5, 10-6, 10-7, etc. by gradient dilution. Each dilution (1mL) was added to a petri dish, mixed well with PDA medium, inverted after the substrate had solidified and incubated at 28 ℃ for 7 d.
And after 7d of culture, selecting a culture dish which completely separates each bacterial colony, selecting bacterial colonies containing a large number of same types in the culture dish, scraping hyphae on the surface of the bacterial colony by using a sterile inoculating ring, inoculating the hyphae on the culture dish containing a PDA culture medium, adding chitosan-silver gels with different concentrations respectively, and uniformly coating the chitosan-silver gels on the PDA culture medium to perform a bacteriostatic experiment. Incubate at 28 ℃ for 7 d. And taking a picture for observation and recording.
All the above aseptic operations were carried out in a clean bench and all the used aseptic vessels and substances were autoclaved at 121 ℃ for 30 min.
Example 2
The silver nitrate stock solution 100 μ L in the above example 1 was taken and added to a 2.0mL screw bottle which was cleaned, dried and protected from light, 800 μ L of the chitosan stock solution which was prepared in the same manner as in example 1 was added rapidly and shaken rapidly, and then 100 μ L of the sodium hydroxide stock solution which was prepared in the same manner as in example 1 and 100 μ L were added and shaken rapidly and shaken uniformly. Clear and transparent chitosan-silver hydrogel can be formed.
Grape preservation experiment steps: fresh grapes are divided into four parts, and the grapes are respectively treated by chitosan-silver sol with the concentration of 6.0 percent, chitosan solution with the same mass concentration and silver solution with the same mass concentration, so that the grapes are not treated to be CK group. And respectively measuring the indexes such as weight loss rate, rotting rate, soluble solid, titratable acid, vitamin C content and the like every day, and photographing and recording.
The preparation process, the main features and the application of the invention have been shown and described above. It will be appreciated by persons skilled in the art that the present invention is not limited by the embodiments described above. The scope of the invention is defined by the appended claims.
Claims (4)
1. The preparation method of the antibacterial complex hydrogel is characterized by comprising the following steps:
dissolving silver nitrate crystal with ultrapure water in a dark place to prepare silver nitrate stock solution;
dissolving chitosan powder with ultrapure water, adding glacial acetic acid to promote the dissolution of chitosan, and preparing a chitosan stock solution;
and (3) taking 100 mu L of silver nitrate stock solution, adding 800 mu L of chitosan stock solution and 100 mu L of sodium hydroxide solution to prepare 1.0mL of mixed solution, and quickly shaking up to form the chitosan-silver hydrogel.
2. The method according to claim 1, wherein the silver nitrate stock solution has a concentration of 0.6mol/L, the chitosan stock solution has a concentration of 5mg/mL, and glacial acetic acid having a volume ratio of 1% is added.
3. The composite hydrogel prepared by the preparation method of claim 1, wherein the chitosan-silver hydrogel is a clear and transparent hydrogel which is obtained by the coordination of chitosan and silver ions and the self-assembly of the chitosan and the silver ions through non-covalent bonds, and has the property of being spreadable.
4. The application of the chitosan-silver hydrogel synthesized by the preparation method of claim 1 in bacteriostasis and preservation of grapes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110386009.0A CN113273605A (en) | 2021-04-09 | 2021-04-09 | Chitosan-silver complex antibacterial gel, preparation method and fresh-keeping application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110386009.0A CN113273605A (en) | 2021-04-09 | 2021-04-09 | Chitosan-silver complex antibacterial gel, preparation method and fresh-keeping application thereof |
Publications (1)
| Publication Number | Publication Date |
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| CN113273605A true CN113273605A (en) | 2021-08-20 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110386009.0A Pending CN113273605A (en) | 2021-04-09 | 2021-04-09 | Chitosan-silver complex antibacterial gel, preparation method and fresh-keeping application thereof |
Country Status (1)
| Country | Link |
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| CN (1) | CN113273605A (en) |
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2021
- 2021-04-09 CN CN202110386009.0A patent/CN113273605A/en active Pending
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